Production of single chain Fab (scFab) fragments in Bacillus megaterium.
Production of single chain Fab (scFab) fragments in Bacillus megaterium.
复制标题
在巨大芽孢杆菌中生产单链 Fab (scFab) 片段。
DOI：
10.1186/1475-2859-6-38
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发表时间：
2007-11-27
影响因子：
6.4
通讯作者：
Duebel, Stefan
中科院分区：
文献类型：
--
作者：
Jordan, Eva;Al-Halabi, Laila;Schirrmann, Thomas;Hust, Michael;Duebel, Stefan
研究、诊断和治疗中对抗原结合试剂的需求对新的抗体形式以及适当的生产系统提出了问题。最近，在革兰氏阴性杆菌中产生了结合单链抗体(ScFv)和Fab片段特性的新型单链抗体形式(ScFab)。在这项研究中，我们评价了革兰氏阳性菌巨型芽孢杆菌对单链抗体和单链抗体的重组生产，并与大肠杆菌进行了比较。在巨大芽孢杆菌和大肠杆菌中产生溶菌酶特异性的D1.3单链抗体。从大肠杆菌周质部分和培养上清液中纯化后的ScFab总产量略高于巨大芽孢杆菌培养上清。然而，通过分析抗原结合活性确定的功能性单链抗体的产量在两个生产系统中是相同的。此外，还在巨型芽孢杆菌中产生了人C反应蛋白特异性的单链抗体片段。巨型芽孢杆菌产生的抗C-反应蛋白单链抗体的总产量略低于大肠杆菌，而纯化的单链抗体的比活力高于大肠杆菌。巨大芽孢杆菌允许分泌产生抗体片段，包括新的单链抗体形式。功能抗体片段的产量和质量与在大肠杆菌中生产的周质相媲美。
The demand on antigen binding reagents in research, diagnostics and therapy raises questions for novel antibody formats as well as appropriate production systems. Recently, the novel single chain Fab (scFab) antibody format combining properties of single chain Fv (scFv) and Fab fragments was produced in the Gram-negative bacterium Escherichia coli. In this study we evaluated the Gram-positive bacterium Bacillus megaterium for the recombinant production of scFab and scFvs in comparison to E. coli. The lysozyme specific D1.3 scFab was produced in B. megaterium and E. coli. The total yield of the scFab after purification obtained from the periplasmic fraction and culture supernatant of E. coli was slightly higher than that obtained from culture supernatant of B. megaterium. However, the yield of functional scFab determined by analyzing the antigen binding activity was equally in both production systems. Furthermore, a scFv fragment with specificity for the human C reactive protein was produced in B. megaterium. The total yield of the anti-CRP scFv produced in B. megaterium was slightly lower compared to E. coli, whereas the specific activity of the purified scFvs produced in B. megaterium was higher compared to E. coli. B. megaterium allows the secretory production of antibody fragments including the novel scFab antibody format. The yield and quality of functional antibody fragment is comparable to the periplasmic production in E. coli.
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影响因子：
12.6
作者：
Kramer, K;Fiedler, M;Hock, B
通讯作者：
Hock, B
影响因子：
3.9
作者：
Schenk, Joerg A.;Sellrie, Frank;Micheel, Burkhard
通讯作者：
Micheel, Burkhard
DOI：
10.1093/protein/13.10.725
发表时间：
2000-10-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Schmiedl, A;Breitling, F;Dübel, S
通讯作者：
Dübel, S
DOI：
10.1016/s0167-4781(03)00038-1
发表时间：
2003-04-15
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
影响因子：
--
作者：
Shiroza, T;Shinozaki-Kuwahara, N;Abiko, Y
通讯作者：
Abiko, Y
影响因子：
3.8
作者：
Chen, C;Snedecor, B;Champion, KM
通讯作者：
Champion, KM